克隆绵羊——多莉（DOLLY）的研究进展

多莉，第一例大型克隆哺乳动物，由一只称为ＦｉｎｎＤｏｒｅｔ的６岁母羊乳腺体细胞的基础生命物质经细胞核转移等操作而产生。成果分布后，持怀疑观点的学者提出，提供体细胞的母羊是否本身处在妊娠状态？推测多莉是由胚胎细胞产生，并非体细胞本身克隆所致。由于６岁的母羊死于１９９５年，只能用其冷冻保存的乳腺组织在Ｈａｎｎａｈ研究中心进行细胞群体的分析。首先，用微卫星扩增技术，以三套引物进行测定；其次，进行ＤＮＡ指     

恒河猴群微卫星DNA多态性的分析

目的　确立一种对恒河猴群个体的遗传物质进行准确可靠、快速简便的遗传检测方法。方法　利用聚合酶链反应 (PCR)扩增技术对 2 0只恒河猴群个体间进行了DNA多态性的分析。结果　筛选出 9个微卫星DNA位点具有显著多态性 ,4个微卫星DNA位点没有多态性 ,还有 2个位点等位基因数目较少。结论　利用这些多态性微卫星位点建立一种对恒河猴群个体进行有效、准备可靠、快捷简便的遗传背景监测方法。     

我国育成小麦品种的遗传多样性演变

对我国小麦育成品种初选核心种质(1680份)的78个微卫星标记(SSR)位点进行了扫描,并就此对50年来育成品种的遗传多样性进行了评价和分析,得到以下结果和结论:(ⅰ)74对SSR荧光引物共检测到1336个等位变异,其中1253个等位变异可以定位在71个位点上.这71个位点上检测到的每个位点等位变异数为4~44个,平均17.6个;多态性信息指数(PIC)为0.19~0.89,平均为0.69.(ⅱ)三个基因组的平均等位变异丰富度为B>A>D,遗传多样性指数为B>D>A.(ⅲ)7个部分同源群的平均等位变异丰富度为2=7>3>4>6>5>1,遗传多样性指数为7>3>2>4>6>5>1.结合两个指标分析,第7部分同源群具有最高的多样性,而1,5群多样性最低.(ⅳ)21条染色体中,7A,3B和2D三条染色体遗传多样性较高,而2A,1B,4D,5D和1D的遗传多样性偏低.(ⅴ)育成品种遗传多样性指数以50年代的最高,以后越来越低,但年代间变化较平缓;品种间平均遗传距离以50年代最高(0.731),以后逐渐减小,各年代依次为0.711,0.706,0.696和0.695.品种遗传基础狭窄化问题日趋突出,应引起有关部门和育种家的关注.     

微卫星DNA聚丙烯酰胺凝胶电泳(PAGE)银染法的改良

为了筛选和检测绵羊种群中具有较好表现的微卫星多态性,采用Touch-down PCR方法扩增绵羊基因组上的微卫星DNA序列后,经聚丙烯酰胺凝胶电泳分离,采用改良的银染法对微卫星位点进行多态性检测,染色结果与常规银染法相比,该方法具有灵敏度高、背景浅、污染小、条带清晰等突出特点,能有效地控制染色背景,显著提高检测分辨率,整个染色过程所需时间短,具有广泛的推广价值。     

运用微卫星DNA标记分析我国野生鹌鹑遗传多样性

运用微卫星DNA标记对我国境内分布的两种野生鹌鹁（野生日本鸣鹁、野生普通鹌鹑）和家鹑群体的遗传多样性进行分析。通过计算反映群体变异的多态信息含量（PIC）、固定指数、平均杂合度、基因分化系数等相关指标,结果表明：野生普通鹌鹑群体遗传多样性更为丰富,每个座位平均检测出4．67个等位基因,群体多态信息含量和平均杂合度亦为最高,分别为0．5732和0．6621;家鹁最低,分别为0．5467和0．5933;野生日本鸣鹑介于两者之间;在3个鹌鹑群体中,野生日本鸣鹑与家鹑群体遗传多样性差异程度较小。以标准遗传距离为基础的模糊聚类分析发现：家鹑与野生日本鸣鹑群体模糊等价矩阵系数为0．937,而与野生普通鹌鹑的系数为0．738,这也表明家鹑与野生日本鸣鹁有更近的亲缘关系,进一步从分子水平上证实家鹑起源于野生日本鸣鹁。     

猪肥胖基因5''''调控区与内含子1的测序分析及其两个新微卫星的发现

根据猪肥胖基因(Obese gene,Ob)部分序列设计引物,并通过Southern杂交和杂交阳性片段的克隆进行测序,首次得到猪Ob基因内含子1和5'调控区16.4 kb序列,将外显子1定位在起始密码子上游11.1 kb处,同时在内含子1中发现了两个新的微卫星,命名为SW200和SW160.调控区潜在的调控元件分析显示,-1～ -300 bp区间包含了C/EBP和两个Sp1,可能更直接高效的调节其转录.为研究SW200和SW160与一些重要经济性状的关系,分析了等位基因和基因型频率,用SAS8.2软件分析显示,两个微卫星多态仅对二花脸猪的头胎总产仔数有显著的效应.     

异源四倍体鲫鲤及其原始亲本遗传变异的微卫星标记分析

采用从鲤中分离出来的32对微卫星DNA标记,对异源四倍体鲫鲤、红鲫和野鲤的基因组DNA进行了研究。在筛选出的15对微卫星引物中,随引物不同,各等位基因数为1～8个,大小在100～420bp之间。从3个不同群体内部的遗传相似系数来看,异源四倍体鲫鲤个体之间的遗传相似系数最大,说明异源四倍体鲫鲤群体内部的遗传变异程度最低,已经形成了一个遗传性状稳定的群体。从3个不同群体之间的遗传相似系数来看,异源四倍体鲫鲤和红鲫遗传相似系数为0．5625,和野鲤的遗传相似系数为0．5125,说明异源四倍体鲫鲤接受原始母本的遗传物质比原始父本野鲤要多一些。微卫星标记与以前报道的RAPD标记的检测结果是相似的,然而由微卫星标记获得的种群内和种群间的遗传距离均大于RAPD,说明微卫星标记比RAPD标记显示出更高的个体多态性。     

中国人胃癌染色体17q21区域基因遗传不稳定性研究

To investigate microsatellite instability (MSI) and loss of heterozygosity (LOH) of locus D17S396, D17S579 and D17S855, and their effect on the expression of nm23H1 and BRCA1 of gastric cancer, which would provide experimental basis for clinical treatment and prognosis analysis of gastric cancer. DNA was extracted from paraffin-embedded materials. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was used to analyze MSI and LOH. Expression of nm23H, and BRCA1 was detected by Envision immuno-histochemistry and Leica-Qwin computer imaging techniques. In the forty cases of gastric cancer, the frequency of MSI, LOH and nm23H1 protein were 20.00%, 17.50% and 55.00% respectively at locus D17S396, while at locus D17S579, the frequency of MSI, LOH and BRCA1 protein were 22.50%, 15.00% and 37.50% respectively; at locus D17S855, the frequency of MSI, LOH and BRCA1 of thirty-seven cases were 18.92%, 18.92%, 37.84% respectively. In tumor node metastasis (TNM) staging, at locus D17S396, D17S579 and D17S855, MSI in stages I + II appeared more frequently than that in stages III + IV, while LOH appeared the contrary tendency. In the group of metastasis of gastric cancer, MSI had a less frequency (5.00%) than that with no metastasis (35.00%, P < 0.05) at locus D17S396, but LOH appeared more frequently (30.00%) than that with no metastasis (5.00%, P < 0.05). At locus D17S579, MSI had an increasing tendency with the degree of tumor differentiation (50.00% in high differentiation cases, 20.00% in middle differentiation cases, and 0% in low differentiation cases, P < 0.05). The frequency of nm23H1 and BRCA1 protein in stages TNM I + II was higher than that in stages TNM III + IV; and that in higher differentiation cases was higher than in poor differentiation cases. The frequency of nm23H1 protein in the group of metastasis (30.00%) was less than that with no metastasis significantly (80.00%, P<0.01). The frequency of nm23H1 protein in the group positive to MSI (87.50%) was higher than that in the group negative to MSI (46.88%, P < 0.05). However, nm23H1 protein in group positive to LOH (14.29%) was lower than that in the group negative to LOH (63.64%, P < 0.05). The frequency of BRCA1 protein in the group positive to MSI (66.67%) was more than that in the group negative to MSI (29.03%, P < 0.05). The results of experiments indicate that MSI and LOH may separately control the development of sporadic colon cancer with different pathways. MSI may be an early period molecule marker for sporadic colon cancer, enhanced expression of nm23H1 protein can effectively inhibit colon cancer metastasis and improve prognosis of sporadic colon cancer patients. By comparison, LOH mostly arises in the late period of sporadic colon cancer and endows a high aggressive and poor prognostic phenotype. nm23H1 protein could effectively restrain gastric cancer metastasis and development; and BRCA1 protein could restain tumor from becoming lower differentiation.     

牙鲆CA/GT微卫星标记的筛选

采用生物素选择杂交法与放射性同位素杂交法相结合的技术,成功地从牙鲆(Paralichthys olivaceus)基因组中分离出含有CA/GT重复类型的微卫星序列。通过两轮淘选,共获得526个阳性菌落。测序其中的119个菌落,结果获得133个含有微卫星座位的序列。除了两个复合型微卫星外(1.5%),完美型63个(47.37%),非完美型68个(51.13%)。设计并合成22对微卫星引物,对8个人工雌核发育家系的亲本进行遗传背景分析。PCR结果表明,4对引物无扩增带或者扩增带不是目的条带,1对引物表现为单态,其余17对引物均呈多态性,平均每个座位产生5.2个复等位基因,杂合度为0.375～0.846,多态信息含量为0.305～0.823。结果表明,所筛选的大部分微卫星标记能够用于牙鲆群体遗传学研究。     

西门塔尔牛产奶性状的微卫星标记遗传效应分析

在4条染色体上选择12个微卫星标记对西门塔尔牛育种核心群6个父系组成的150头母牛产奶性状(包括乳脂、乳蛋白、乳糖、干物质含量和奶中体细胞数)进行分子标记遗传效应分析.结果表明:12个微卫星位点都具有高度多态性,杂合度(H)均在0.64～0.86之间,多态信息含量(PIC)达0.60以上,最高为0.85(ILST093);位点ILST093对奶中体细胞数有显著性影响(P＜0.05);BMS711对乳脂率有显著性影响(P＜0.05);BM1905与奶中乳糖含量呈显著相关(P＜0.05);BM6438与5个产奶性状均无相关性.